Malaysian Journal of Analytical Sciences Vol 20 No 1
(2016): 31 - 43
SPECTROSCOPIC
ANALYSIS OF RHAMNOLIPID PRODUCED BY PRODUCED BY Pseudomonas aeruginosa UKMP14T
(Analisis Spektroskopik Ramnolipid yang Dihasilkan oleh P. aeruginosa UKMP14T)
Noramiza Sabturani1,
Jalifah Latif2, Shahidan Radiman3, Ainon Hamzah1*
1School of Biosciences and Biotechnology
2School of Chemical Sciences and Food Technology
3School of Applied Physics
Faculty of Science
and Technology,
Universiti
Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia
*Corresponding author: ainonh@gmail.com
Received:
21 September 2015; Accepted: 20 November 2015
Abstract
Biosurfactant produced by Pseudomonas aeruginosa UKMP14T was
optimized by growing the isolate in mineral
salt medium (MSM) supplemented with 1% (v/v) glycerol and 1.3 g/L ammonium
sulphate with C/N ratio of 14:1. The culture medium was incubated at 37°C, with
an agitation speed of 150 rpm for 7 days. P. aeruginosa UKMP14T produced
biosurfactant at 0.8 g/L after 7 days incubation. Anthrone assay proved
biosurfactant was glycolipid. The biosurfactant was characterized by scanning
electron microscope-energy dispersive X-ray spectroscopy (SEM-EDX) in addition
to Fourier transform infra-red (FT-IR), nuclear magnetic resonance (NMR) and
electrospray ionization-mass spectrometry (ESI-MS). The SEM-EDX analysis
indicated the presence of carbon and oxygen elements by 78% and 22% (atomic %),
respectively. FT-IR absorption spectra indicated the functional groups of
rhamnolipid were located at 3308.46, 2922.91, 2857.09 and 1730.96 cm-1.
ESI-MS/MS analyses identified P.
aeruginosa UKMP14T produced rhamnolipid with two fatty acids-nine
congeners, L-rhamnosyl-L-rhamnosyl-β-hydroxydecanoyl-β-hydroxydecanoate
(Rha-Rha-C10-C10) (m/z
649) which formed the main compound. The critical micelle concentration
(CMC) of this rhamnolipid was established at 30 mg/L (32 dynes/cm). The
characteristics of biosurfactant produced by P. aeruginosa UKMP14T indicated it has a high potential in industrial
dan bioremediation application.
Keywords: biosurfactant, Pseudomonas aeruginosa, rhamnolipid,
spectroscopic analysis, CMC value
Abstrak
Biosurfaktan dihasilkan oleh Pseudomonas aeruginosa UKMP14T
dioptimumkan dalam medium garam mineral (MSM) yang ditambahkan dengan 1% (i/i)
gliserol dan 1.3 g/L ammonium sulfat dengan nisbah C/N iaitu 14:1. Medium
kultur ini dieram pada 37°C, dengan kelajuan goncangan pada 150 psm selama 7
hari. P. aeruginosa UKMP14T
menghasilkan biosurfaktan sebanyak 0.8 g/L selepas 7 hari pengeraman. Asai
antron membuktikan biosurfaktan terhasil adalah glikolipid. Biosurfaktan
tersebut dicirikan oleh mikroskop pengimbasan elektron-spektroskopi tenaga
serakan sinaran-X (SEM-EDX), dengan penambahan Infra-merah transformasi Fourier
(FT-IR), resonan magnetik nuklear (NMR) daknn spektrometri jisim-pengionan
elektrosemburan (ESI-MS). Analisis SEM-EDX menunjukkan kehadiran unsur karbon
dan oksigen masing-masing sebanyak 78% dan 22% (% atomik). Spektrum penyerapan
FT-IR menunjukkan kumpulan berfungsi ramnolipid pada 3308.46, 2922.91, 2857.09
dan 1730.96 cm-1. Analisis ESI-MS/MS mengenalpasti P. aeruginosa UKMP14T menghasilkan
ramnolipid dengan sembilan kongener-dua asid lemak,
L-ramnosil-L-ramnosil-β-hidroksidekanoil-β-hidroksidekanoat (Rha-Rha-C10-C10)
(m/z 649) sebagai sebatian utama.
Kepekatan misel kritikal (CMC) ramnolipid ini dikesan pada 30 mg/L (32
dynes/cm). Ciri-ciri biosurfaktan yang dihasilkan oleh P. aeruginosa UKMP14T menunjukkan ia mempunyai potensi tinggi untuk
di aplikasi dalam industri dan bioremediasi.
Kata kunci: biosurfaktan, Pseudomonas aeruginosa, ramnolipid,
analisis spektroskopik, nilai CMC
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